STEM CELL CORE 1. MAIN OBJECTIVES AND NEW DIRECTIONS The Stem Cell Core will be a new facility. Currently, there is no unified Stem Cell Core Facility to link neuroscientists on the La Jolla Torrey Pines Mesa. Therefore, this Stem Cell facility will constitute a new Core to foster stem cell projects and collaborations among neuroscientists in the San Diego area. Interest in human embryonic stem cells (hESCs) has grown steadily since the report of the first hESC line in 1998. hESCs are attractive because of their potential as cell therapy, either directly replacing cells or by using them as vehicles for gene therapy (Muller et al. 2006). But the cells themselves are intrinsically interesting, because of their unique qualities of pluripotence and self-renewal, and these qualities have led to a resurgence of interest in fundamental cell biology. hESC are the perfect tools to study events that occur during human neurogenesis, both to understand normal development and to identify what goes wrong during neurological disease. The versatility of hESC means that from the same population one can generate multiple cell types that can then be used to dissect complex cellcell interactions. For example, by differentiating hES cells into motor neurons, skeletal muscle cells, and astroglia, one can do mix-and-match experiments to identify the vital components of degeneration in ALS. By generating human/mouse chimeric brains using transplanted human neural stem cells or neuronal precursors derived from hESC, it will be possible to ask how human neurons react to the amyloid plaques that develop in transgenic models of Alzheimer's disease. Additionally, hESCs will eventually be used for transplantation therapies in regenerative medicine. This Core facility proposes to use only approved Presidential ES cell lines. The funds requested are particularly important because Prop 71 which was to fund stem cell work in California has not become a reality, and it is currently not clear if these funds will become available in the future. In addition to hESCs, this core will allow neuroscientists on the La Jolla Torrey Pines Mesa access to methods regarding the use of murine (m)ES cells. Additionally, the Stem Cell Core has expertise with neural progenitor cells (NPCs) derived both from hES and mES cells. Moreover, NPCs from fetal human and mouse brains (obtained by workers at BIMR and UCSD), and adult rodent NPCs (from the laboratory of Fred Gage at The Salk Institute) will be available through this Stem Cell Core. The origins of all of these cells and cell lines have received both Institutional approval and Presidential approval, where necessary for hESC lines. The purpose of the Stem Cell Core will be to provide training and fundamental analysis tools for stem cell research throughout the La Jolla Torrey Pines Mesa. Currently, while there is an NIH-sponsored Stem Cell Center at the Burnham Institute for Medical Research (BIMR), it is aimed at specific projects and is insufficient to provide support for local neuroscientists who would like to use it. With this grant, we propose to take advantage of the BIMR's experienced Stem Cell Center faculty and staff and expand the capacity of the core laboratory to make the facilities available to all neuroscientists in the La Jolla Torrey Pines Mesa area (UCSD, Salk, BIMR and TSRI). The Stem Cell Core will be directed by Dr. Alexey Terskikh (BIMR, who trained with Prof. Irving Weismann, a well-known stem cell expert at Stanford. Co-directors of the facility will be stem cell experts Drs. Evan Snyder and Jeanne Loring (at BIMR), and Fred (Rusty) Gage and Juan Carlos Belmonte at The Salk Institute. The overall objective of the Stem Cell core is to lower the barrier for neuroscientists to involve stem cells in their research projects, by providing training and expert advice from experienced stem cell biologists. This will improve communication among scientists in a variety of neuroscience subspecialties to foster interdisciplinary collaborations and novel approaches. By sharing common techniques and accumulated knowledge about the fundamental biology of human ES cells and somatic (adult) stem cells, there will be a considerable savings in time, effort, and cost. The Stem Cell Core will play an important role in ensuring that the users are kept up to date with the standardization of hESC methods called for by the international stem cell scientific community (Andrews et al. 2005;Loring 2005). The principal focus of this core is to provide stem cell-specific technologies that will lead to new insights in neuroscience and aid in development of clinical applications for stem cells. For example, correlation of the characteristics of stem cells in culture and their behavior after transplantation will help in decisions about which cells to use and what quality control measures will be necessary for clinical use. Core support will include culturing of ES and other stem cells, karyotyping, SNP genotyping, microarray analysis, immunohistochemistry for stem cell and differentiation markers, assays for pluripotence and differentiation, and access to BIMR's extensive databases of stem cell information. In addition, the stem cell core faculty will offer advice for experimental design, data analysis, interpretation of results, and assistance with manuscripts and grant applications. Stem cell core faculty will also serve as mentors for postdocs and young faculty members, to help them publish and obtain grant support.